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研究生:張育誠
研究生(外文):Yu-Cheng Chang
論文名稱:台灣黑腹絨鼠族群分化之研究
論文名稱(外文):Population Divergence of Père David’s Red–backed Vole (Eothenomys melanogaster) in Taiwan
指導教授:林良恭林良恭引用關係
指導教授(外文):Liang-Kong Lin
學位類別:碩士
校院名稱:東海大學
系所名稱:生命科學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:73
中文關鍵詞:黑腹絨鼠分化地理親緣DNA 序列核型
外文關鍵詞:Eothenomys melanogasterdivergencePhylogeographyDNA sequenceKaryotype
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台灣的黑腹絨鼠(Eothenomys melanogaster)棲息於中、高海拔山區,屬較適冷且為營地道生活之田鼠類。細胞遺傳研究發現,台灣島內的黑腹絨鼠有兩種形態的X染色體存在,顯示島內族群可能已有相當程度的分化。為瞭解分化的程度與探討其可能原因,本研究持續進行染色體核型研究,並以3段遺傳標記,探討臺灣黑腹絨鼠各地理族群間之分化現象。結果顯示,X染色體核型、粒線體Cyt b完整序列(1143 bp)、核基因IRBP exon 1部份序列(1260 bp)與性聯遺傳基因G6pd intron 1 部分序列(579 bp)的親緣關係分析皆支持台灣黑腹絨鼠南、北兩大地理群以及與大陸四川黑腹絨鼠族群的關係。其中,北群的X染色體為中節端位(Acrocentric),南群的為次中節中位(Submetacentric)。北群與南群的Cyt b遺傳距離相差3.7%;北群與南群的IRBP序列具有6個可區別位點(diagnostic site);北群與南群的G6pd序列具有2個可區別位點。依分子時鐘估算兩群之分歧時間約為0.59 百萬年前(Mya),推測台灣黑腹絨鼠於Cromerian間冰期(0.63-0.56 Mya)時氣候變遷,造成台灣南、北地理族群遭受隔離而分化。由單型網絡關係推測南北族群間冰期避難所可能位於阿里山與觀霧兩處。此外,台灣黑腹絨鼠南、北群間之遺傳差異,就其遺傳距離3.7%(Cyt b)與核型差異來看,應可視為不同亞種;而台灣黑腹絨鼠與大陸四川黑腹絨鼠之遺傳距離達7.0%(Cyt b),應可將其視為不同種。
Père David’s red–backed vole (Eothenomys melanogaster) inhabits in the middle and high elevations of Taiwan. It is adapted to cold and lives underground in a tunnel system. Cytogenetic studies separate E. melanogaster into two races based on the differently shaped X chromosomes. Populations of E. melanogaster in Taiwan may have deeply diverged genetically. To test the interglacial refugia hypothesis and identify the pattern of phylogeographic differentiation, I investigated variation among the geographical populations of E. melanogaster in Taiwan. Chromosomal analysis, complete mitochondrial cytochrome b sequences (1143 bp), partial nuclear IRBP exon 1 sequences (1260 bp), and complete X chromosome-link gene G6pd intron 1 sequences (579 bp) supported division of E. melanogaster in Taiwan into Northern and Southern groups. The X chromosome of the Northern group was acrocentric, but it was submetacentric in the Southern group. The Cyt b genetic distance (Juke-Cantor model) between Northern and Southern groups was 3.7%. There were 6 diagnostic sites on IRBP and 2 diagnostic sites on G6pd separating Northern and Southern groups. Divergence time between two groups was 0.59 million years ago (Mya) according to mean rate 6.02% per million years (Cyt b). The populations of E. melanogaster in Taiwan were probably restricted to isolated refugia around the Cromerian interglacial period (0.63-0.56 Mya) and then differentiated genetically. The haplotype network indicated Cromerian interglacial refugia around the Alishan area for Southern group and around the Guanwu area for Northern group. Based on the genetic distance, I recommend that the Northern and Southern groups be considered two subspecies. This is because the middle degree of genetic distance and the extremely low gene flow. Since the mean Cyt b genetic distance between E. melanoagaster in Taiwan and Sichuan was greater than the minimal genetic Cyt b distance and almost reached mean Cyt b genetic distance within subgenus. I recommend E. melanogaster in Taiwan and Sichuan be considered two species.
中文摘要 1
英文摘要 3
一、前言 5
二、材料與方法 9
(一)樣本採集 9
(二)實驗方法 9
1. 核型製備 9
2. 去氧核糖核酸分析 11
(1)DNA萃取 11
(2)聚合酶鏈反應及去氧核糖核酸定序 11
A. 粒線體DNA細胞色素b序列 12
B. 核DNA視網膜感光細胞間視黃醇類結合蛋白exon 1部分序列 13
C. X性染色體連鎖基因葡萄糖六磷酸脫氫酶intron 1部分序列 14
(3)序列處理及親緣遺傳分析 15
三、結果 18
(一)核型分析 18
1. 台灣黑腹絨鼠野外族群 18
(1)傳統染色法核型 18
(2)Giemsa條帶染色法核型 19
(3)Constitutive heterochromatin條帶染色法核型 19
(二)分子資料分析 20
1. 粒線體DNA細胞色素b序列 20
(1)DNA序列分析 20
(2)親緣遺傳分析 21
2. 核DNA視網膜感光細胞間視黃醇類結合蛋白exon 1部分序列(IRBP) 24
(1)DNA序列分析 24
(2)親緣遺傳分析 24
(3)台灣黑腹絨鼠野外族群疑似雜交個體 26
3. X性染色體連鎖基因葡萄糖六磷酸脫氫酶intron 1部分序列(G6pd) 26
(1)DNA序列分析 26
(2)親緣遺傳分析 27
四、討論 29
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